An ex vivo perfused ventilated murine lung model suggests lack of acute pulmonary toxicity of the potential novel anticancer agent (−)-englerin A

Abstract

Abstract(−)-Englerin A (EA), a potential novel anti-cancer drug, is a potent selective activator of classical transient receptor potential 4 and 5 (TRPC4, TRPC5) channels. As TRPC4 channels are expressed and functional in the lung endothelium, possible side effects such as lung edema formation may arise during its administration. Well-established in vivo rodent models for toxicological testing, however, rapidly degrade this compound to its inactive derivative, englerin B. Therefore, we chose an ex vivo isolated perfused and ventilated murine lung (IPVML) model to detect edema formation due to toxicants, which also reduces the number of incriminating animal experiments required. To evaluate the sensitivity of the IPVML model, short-time (10 min) drops of the pH from 7.4 down to 4.0 were applied, which resulted in linear changes of tidal volumes, wet-to-dry weight ratios and incorporation of FITC-coupled dextran particles from the perfusate. As expected, biological activity of EA was preserved after perfusion in the IPVML model. Concentrations of 50–100 nM EA continuously perfused through the IPVML model did not change tidal volumes and lung weights significantly. Wet-to-dry weight ratios were increased after perfusion of 100 nM EA but permeation of FITC-coupled dextran particles from the perfusate to the lung tissues was not significantly different. Therefore, EA shows little or no significant acute pulmonary toxicity after application of doses expected to activate target ion channels and the IPVML is a sensitive powerful ex vivo model for evaluating acute lung toxicity in accordance with the 3R rules for animal experimentation.

Transient Receptor Potential (TRP) Channels in Airway Toxicity and Disease: An Update

“…During lung transplantation, I/R-induced injury is a major reason for graft failure [ 158 ]. Next to toxicant-induced edema [ 159 ], I/R-induced edema can be mimicked in the isolated perfused and ventilated mouse lung (IPVML) [ 160 ]. Most interestingly, I/R-induced edema was absent in TRPC6-deficient mice, and edema formation in double TRPC6/TRPV4−/− lungs was similar to WT mice [ 119 ], which may be explained by an antagonistic function of TRPC6 and TRPV4 in the lung endothelium and epithelium, respectively (reviewed in [ 8 ]).…”